title: Label-free detection of single-nucleotide polymorphisms associated with myeloid differentiation-2 using a nanostructured biosensor.
abstract: House dust mites are the major source of indoor allergens that are responsible for asthma. The major dust mite allergen is the group II allergen, Der p2. Myeloid differentiation-2 (MD-2) acts as an essential component in the CD14-TLR4 (toll-like receptor)/MD-2 receptor complex for Der p2 recognition. Mutations of the MD-2 associated gene on chromosome 8 degrade a human's innate responses. In this study, we report the effective detection of mutations to the MD-2 gene promoter, using a sensitive nanostructured biosensor with a sensing electrode of gold nanoparticles (GNPs) uniformly deposited in a nanohemisphere array. The 70 mer MD-2 expressed gene fragment was used to probe gene mutation. The complementary target, containing 156 mer nucleotide, was prepared using the fresh blood from patients with allergic disease. A total of 37 target samples, including 19 gene mutated samples and 18 normal samples, were prepared by a 20 cycles PCR process, and used for discrimination experiments. Experimental results illustrated that the nanostructured biosensor clearly discriminates between mutated and non-mutated MD-2 allergy genes.
authors.affiliation: Department of Mechanical Engineering, National Chung-Hsing University, Taichung 40227, Taiwan.
authors.forename: Yu-Ting
authors.initials: YT
authors.lastname: Chin
authors.forename: En-Chih
authors.initials: EC
authors.lastname: Liao
authors.forename: Chia-Che
authors.initials: CC
authors.lastname: Wu
authors.forename: Gou-Jen
authors.initials: GJ
authors.lastname: Wang
authors.forename: Jaw-Ji
authors.initials: JJ
authors.lastname: Tsai
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D058266:Dielectric Spectroscopy / Q000379:methods; D006801:Humans; D051218:Lymphocyte Antigen 96 / Q000235:genetics*; D009154:Mutation; D049329:Nanostructures / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic
chemical_list: C495377:LY96 protein, human; D051218:Lymphocyte Antigen 96
keywords: Electrochemical impedance spectroscopy (EIS) analysis; Myeloid differentiation-2 (MD-2) gene promoter; Nanostructured biosensor; Single nucleotide polymorphism (SNP) detection